Summary
An antigen common to purported centriolar and basal body regions of a variety of cell types was previously visualized by immuno-fluorescence microscopy. The present study demonstrates the localization of the antigen relative to the defined basal body structures of ciliated tracheal cells at the electron-microscopic level. After ethyldimethylaminopropyl carbodiimide-glutaraldehyde-saponin (EGS) fixation and permeabilization, immunoferritin labeling is consistently found associated with amorphous electron-opaque material in proximity to basal bodies and their ciliary rootlets, but not with basal body microtubules themselves. This distribution pattern is distinct from that of other proteins found in the apical region of ciliated cells, such as calmodulin. It is proposed that the dense material may be analogous to pericentriolar material of centrosomes.
Similar content being viewed by others
References
Albrecht-Buehler G, Bushnell A (1980) The ultrastructure of primary cilia in quiescent 3T3 cells. Exp Cell Res 126:427–437
Berns MW, Meredith Richardson S (1977) Continuation of mitosis after selective laser microbeam destruction of the centriolar region. J Cell Biol 75:977–982
Brenner S, Branch A, Meredith S, Berns MW (1977) The absence of centrioles from spindle poles of rat kangaroo (PtK2) cells undergoing meiotic-like reduction division in vitro. J Cell Biol 72:368–379
Buckley IK, Porter KR (1967) Cytoplasmic fibrils in living cultured cells: a light and electron microscope study. Protoplasma 64:349–380
Connolly JA, Kalnins VI (1978) Visualization of centrioles and basal bodies by fluorescent staining with nonimmune rabbit sera. J Cell Biol 79:526–532
Fonte VG, Searls RL, Hilfer SR (1971) The relationship of cilia with cell division and differentiation. J Cell Biol 49:226–229
Fulton C (1971) Centrioles. In: Reinert J, Ursprung H (eds) Origin and continuity of cell organelles. Springer-Verlag, New York, pp 170–221
Gordon RE, Williams KB, Puszkin S (1982) Immune localization of calmodulin in the ciliated cells of hamster trachea epithelium. J Cell Biol 95:57–63
Gould RR, Borisy GG (1977) The pericentriolar material in Chinese hamster ovary cells nucleates microtubule formation. J Cell Biol 73:601–615
Greenwood FC, Hunter WM, Clover JS (1963) The preparation of 131I-labelled human growth hormone of high specific radioactivity. Biochem J 89:114–123
Kasamatsu H, Nehorayan A (1979) Intracellular localization of viral polypeptides during simian virus 40 infection. J Virol 32:648–660
Kasamatsu H, Shyamala M, Lin W (1980) Host antigens in the centriolar region are induced in SV40-infected TC7 cells: SV40 small-t function requirement. Cold Spring Harbor Symp Quant Biol 44:243–252
Lin W, Fung B, Shyamala M, Kasamatsu H (1981) Identification of antigenically related polypeptides at centrioles and basal bodies. Proc Natl Acad Sci USA 78:2373–2377.
Maunoury R (1978) Localisation immunocytochimique de la centrosphere de cellules tumorales humaines par utilisation d'anticorps naturels de lapin. CR Acad Sc Paris, t. 286, Serie D: 503–506
Nenci I, Marchetti E (1978) Concerning the localization of steroids in centrioles and basal bodies by immunofluorescence. J Cell Biol 76:255–260
Ohtsuki I, Manzi RM, Palade GE, Jamieson JD (1978) Entry of macromolecular tracers into cells fixed with low concentrations of aldehydes. Biol Cell 31:119–126
Osborn M, Weber K (1976) Cytoplasmic microtubules in tissue culture cells appear to grow from an organizing center towards the plasma membrane. Proc Natl Acad Sci USA 73:867–871
Raff EC (1979) The control of microtubule assembly in vivo. Int Rev Cytol 59:1–96
Robbins E, Jentzsch G, Micali A (1968) The centriole cycle in synchronized HeLa cells. J Cell Biol 36:329–339
Sandoz D, Gounon P, Karsenti E, Boisvieux-Ulrich E, Laine M-C, Paulin D (1983) Organization of intermediate filaments in ciliated cells from quail oviduct. J Submicrosc Cytol 15:323–326
Scherft JP, Daems WT (1967) Single cilia in chondrocytes. J Ultrastruct Res 19:546–555
Segarini P, Shyamala M, Atcheson CL, Kasamatsu H (1983) The centriolar antigen expression in TC7 cells is dependent on growth conditions and occurs at a particular time point in G1. J Cell Physiol 116:311–321
Sorokin SP (1968) Reconstructions of centriole formation and ciliogenesis in mammalian lungs. J Cell Sci 3:207–230
Steinman RM (1968) An electron microscopic study of ciliogenesis in developing epidermis and trachea in the embryo of Xenopus Laevis. Am J Anat 122:19–56
Tucker RW, Pardee AB, Fujiwara K (1979) Centriole ciliation is related to quiescence and DNA synthesis in 3T3 cells. Cell 17:527–535
Turksen K, Aubin JE, Kalnins VI (1982) Identification of a centriole-associated protein by antibodies present in normal rabbit sera. Nature 298:763–765
Wheatley DN (1969) Cilia in cell-cultured fibroblasts: 1. On their occurrence and relative frequencies in primary cultures and established cell lines. J Anat 105:351–362
Willingham MC (1980) Electron microscopic immunocytochemical localization of intracellular antigens in cultured cells: the EGS and ferritin bridge procedures. Histochem J 12:419–434
Willingham MC, Yamada SS (1979) Development of a new primary fixative for electron microscopic immunocytochemical localization of intracellular antigens in cultured cells. J Histochem Cytochem 27:947–960
Willingham MC, Jay G, Pastan I (1979) Localization of the ASV src gene product to the plasma membrane of transformed cells by electron microscopic immunocytochemistry. Cell 18:125–134
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Fung, B.P., Kasamatsu, H. Immuno-electron-microscopic localization of a centriole-related antigen in ciliated cells. Cell Tissue Res. 239, 43–50 (1985). https://doi.org/10.1007/BF00214901
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00214901